Temporal immersion culture is an in vitro method in which cells, tissues, organs, plants embryos, plantlets or plants are grown by alternate cycles of temporal immersion in liquid culture media and appropriate aeration.
One of the temporal immersion culture methods described is the RITA® system (Etienne and Berthouly, Plant cell, Tissue and Organ Culture, 69:215 (2002)) which consist of an apparatus with a superior compartment vertically adjacent to an inferior compartment, wherein the superior compartment holds e.g., growing plants, and the inferior compartment holds culture media, wherein the culture media migrates temporarily to the superior compartment by the action of compressed air, and wherein the culture media migrates back to the lower compartment by gravity when the compressed air is turned off. This system has many parts and external connections, has a small surface area for culture, and forms bubbles and a humidity gradient.
Another temporal immersion culture system described in the literature consist of two separated compartments, one for e.g., plants and the other for culture media, wherein the culture media migrates from one compartment to another by the action of a pump (Espinosa, Inform. Syst. Biotech. New Reports, 2:7 (2002)). This system is big and hard to handle.
There are another temporal immersion system wherein a cylindrical container with e.g., growing plants, is rotated such that the plants rotating inside the rolling container get submerged in the culture media that lays in the inferior portion of the cylindrical container. This system has the limitation of shearing of the culturing elements that could be delicate and therefore damaged, e.g., a delicate layer of cells that is being cultivated.
Other temporal immersion systems consist of a container with horizontally adjacent compartments, wherein one compartment contains the elements to be cultured, e.g., plants or cells, and another compartment has the culture media, and wherein culture media migrates back and forth from one compartment to the other compartment by a rocking mechanism which causes the culture media to move from the temporarily upper compartment to the temporarily lower compartment because of gravity. These systems create a current from one compartment to the other compartment that is non-homogeneous and which may also cause shearing of the elements being cultivated. Examples of these systems are described in Publication No. MX PA04003837 A (Mexican Patent Application by Gus Petinovich, Leonardo, (2005)), U.S. Pat. No. 6,544,788 B2 by Singh, Vijay (2003), U.S. Pat. No. 6,190,913 B1 by Singh Vijay (2001), and U.S. Pat. No. 6,566,126 B2 by Cadwell, John (2003).
The present invention provides a versatile apparatus that overcomes the limitations of the described systems, wherein the present invention apparatus provides a system in which the cultured elements may be kept horizontally immobilized in an adequate sized surface, and wherein the immersion of the culture cells is made from the bottom to the top gently and homogeneous with appropriate aeration, without shearing nor bubbles formation, and without the use of air pressure nor pumps. The apparatus of the present invention can achieve the culture media immersion of the cultivated elements without any rocking. However, if rocking is desired after the immersion, the apparatus of the present invention may also be adapted for gently rocking.